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[The role of immune cells and inflammatory mediators in pathogenesis of intestinal inflammation].

The initiators of intestinal inflammation are greatly variable, but the mechanisms underlying the immunologically mediated mucosal damage are similar. A great progress in our understanding of the functions of gut-associated lymphoid tissue was achieved because of the advances in immuno-biology and molecular immunology. At the beginning of an inflammation microorganisms and hither to derived products or allergen firstly stimulate and activate the specific and nonspecific immune cells, and then intestine epithelials, macrophages and lymphocytes release various cytokines. These cytokines are able to recruit more immune cells to be activated and more cytokines to be released. A "cascade" is then generated between epithelials, macrophages and lymphocytes. Finally the inflammation in certain area of intestinal tract is generated. The balance between inflammatory factors and anti-inflammatory molecules depends the results of the inflammation. Therefore, the better understanding of the mechanisms on inflammatory and anti-inflammatory factors is helpful for the diagnosis and treatment of intestinal inflammation.

Animals↗

Bacterial infections in cirrhosis.

Spontaneous bacterial peritonitis, urinary tract infections, respiratory infections and bacteremia are the most frequent infective complications in cirrhosis. These infections are due to the concomitant presence of different facilitating mechanisms including changes in the intestinal flora and in the intestinal barrier, depression of activity of the reticuloendothelial system, decreased opsonic activity of the ascitic fluid, neutrophil leukocyte dysfunction and iatrogenic factors among others. The fact, that the probability of having a microorganism responsible for the infection quinolone resistant is higher than 30% should be taken into account when treating any infection in a cirrhotic patient receiving selective intestinal decontamination with quinolones, and therefore, quinolones as empiric treatment are not indicated.

Bacterial Infections↗

Intestinal microflora in 45 crows in Ueno Zoo and the in vitro susceptibilities of 29 Escherichia coli isolates to 14 antimicrobial agents.

Microorganisms from 45 jungle crows (Corvus macrorhynchos) captured from July to December 2002 at Ueno Zoo, Tokyo were identified as Escherichia coli, Proteus mirabilis, Klebsiella pneumoniae, Klebsiella oxytoca, Enterobacter aerogenes, Enterobacter cloacae, Enterobacter agglomerans, Pseudomonas maltophila, Staphylococcus spp., Micrococcus spp., and Streptococcus spp. E. coli showed the highest rate of isolation (21.6%). In an in vitro susceptibility test for 29 isolates of E. coli to 14 antimicrobial agents, all the isolates were resistant to penicillin G, vancomycin, erythromycin, lincomycin, bicozamycin, sulfadimethoxine, and olaquindox. Several isolates of them were also resistant to tetracycline, oxytetracycline, streptomycin, chloramphenicol, and ampicillin. Twenty-nine isolates were divided into 19 serogroups and the most frequently identified serogroups were O8, O114 and O144, which showed the same multidrug-resistant patterns.

Animals↗

[Correction of intestinal microflora in chemotherapeutic dysbacteriosis using bifidobacterial and lactobacterial autologous strains].

The oral administration of kanamycin (40 mg/kg) or ampiox (500 mg/kg) to guinea pigs for 5 days led to disturbances in their normal intestinal microflora, manifested by a sharp decrease in the levels of lactobacteria and bifidobacteria, as well as by the appearance of large amounts of enterobacteria and enterococci, normally not detected in the proximal and distal sections of the intestinal tract. In adult volunteers receiving kanamycin orally in a dose of 40 mg/kg for 5 days disturbances in microbiocenosis also occurred: the amount of enterococci, staphylococci, lactobacteria and bifidobacteria considerably decreased, enterobacteria becoming the dominating microorganisms. Three oral administrations of bifidobacterial and lactobacterial autostrains immediately after the abolition of the antibiotic facilitated the rapid and effective restoration of the intestinal microflora.

Animals↗

Rapid appearance of M cells after microbial challenge is restricted at the periphery of the follicle-associated epithelium of Peyer's patch.

M cells within the follicle-associated epithelium (FAE) of the gut play a central role in the initiation of mucosal immune responses by transporting antigens to the intestinal lymphoid tissue. We have previously demonstrated that the instillation into the gut of a nonenteric microorganism, Streptococcus pneumoniae R36a, is an excellent experimental model to investigate the highly dynamic nature of the FAE in response to microbial challenge. In the present study, S. pneumoniae was introduced into rabbit ileal loops, each one containing a Peyer's patch (PP), and the number of M cells was assessed by morphological and functional characteristics in different areas of the FAE after a short time (1-3 hours). We report that a marked increase in the number of M cells was detected in the periphery, but not in the apical area, of the FAE as early as 1 hour after exposure to S. pneumoniae. Furthermore, a variant of this experiment enabled us to establish that the increased numbers of M cells led to an improved capability of the FAE to transport latex fluorescent microspheres (0.5 microm), highly specific to rabbit M cells, from the gut lumen to the intestinal lymphatic system. In these animals the cisterna chyli was cannulated, and the microparticles were introduced into the intestinal loops after stimulation with pneumococci. The microparticles reaching the lymph were then counted by flow cytometer. We interpreted these results as showing that only enterocytes located within the periphery of the FAE are converted to fully operational M cells by certain microbial interaction and the ability of enterocytes to undergo this conversion may depend on their stage of differentiation.

Animals↗

Impact of antimicrobial agents on human intestinal microflora.

The most common and significant cause of disturbances in the normal intestinal microflora is the administration of antimicrobial agents. The microflora can be influenced by antimicrobial agents because of incomplete absorption of any orally administered antimicrobial agent, secretion of an antimicrobial agent in the bile, or secretion from the intestinal mucosa. In most cases, the influence is not beneficial to the patient because suppression of the indigenous microorganisms often permits potential pathogens to overgrow and cause septic conditions, stomatitis, diarrhea, or colitis. Antimicrobial agents that influence the normal microflora also promote the emergence of antimicrobial-resistant strains. During the last fifteen years, the impact of different antimicrobial agents on the human microflora has been studied by several investigators. In this article published data on the impact of beta-lactam antibiotics, macrolides, tetracyclines, nitroimidazoles, clindamycin and quinolones on the human intestinal microflora are reviewed.

4-Quinolones↗

Lack of effect of orally administered human serum immunoglobulin on the normal human oral and intestinal microflora.

The aim of this study was to investigate the influence of large doses of orally administered human IgG on the normal gastrointestinal microflora of healthy volunteers since human immunoglobulin has been tried as oral prophylaxis and therapy in gastrointestinal infections. Ten adult healthy volunteers received 10 g of IgG orally, once daily for three consecutive days. Aerobic and anaerobic microorganisms were identified in the saliva and stool specimens, using morphological, biochemical and serological tests and gas-liquid chromatography. Although the immunoglobulin preparation contains antibodies against a variety of microorganisms, there were no significant changes in the numbers of different aerobic and anaerobic microorganisms due to the oral intake of the immunoglobulin. IgG may, therefore, be used against pathogens without disturbing the normal oral and intestinal microflora.

Administration, Oral↗

[The frequency of staphylococcal colonization of the intestines in children with the manifestations of dysbacteriosis].

In 2100 children of different age groups the microbiocenosis of the large intestine was studied. The study revealed that the colonization of the mucous membrane of the large intestine with staphylococci developed in 30% of children with intestinal dysbacteriosis. Young children were mainly affected (91%). The prevailing species among isolated staphylococci was S. aureus (86%), capable of persistence in the intestine (30.9%). In children non typing S. aureus strains mainly circulated (70%), and among phage-typing strains isolates of phage group III prevailed (70.2%). The colonization of the intestine with coagulase-negative staphylococci was possible (14%). Microecological intestinal disturbances in children of different age groups were characterized by different degrees of changes in normal microflora with the prevalence of opportunistic microorganisms in the microbial picture.

Bacteriophage Typing↗

Influence of the interdigestive myoelectric complex on enteric flora in the rat.

This study was designed to define the role of the interdigestive myoelectric complex in small intestinal bacteriostasis. In rats, six monopolar electrodes were surgically sewn to the small intestine at equal intervals. One week later myoelectric activity was recorded. Under different experimental conditions, segments of duodenum and ileum were cultured quantitatively, both aerobically and anaerobically. Five groups of 6 electrode-equipped animals each were studied after an overnight fast: rats in which (a) the interdigestive myoelectric complex was present, (b) the interdigestive myoelectric complex was disrupted for 6 h using morphine sulfate, (c) the interdigestive myoelectric complex was disrupted for 15 h using morphine sulfate, (d) the interdigestive myoelectric complex was disrupted for 15 h using phenylephrine, and (e) the interdigestive myoelectric complex returned after 15 h of morphine sulfate effect. In control rats and during baseline records before drug administration in the other four groups, the interdigestive myoelectric complex was present. Activity fronts cycled at regular intervals in the proximal small intestine and moved aborally. Activity fronts disappeared following both morphine and phenylephrine, with varying degrees of inhibition of spike activity. Titers of microorganisms increased after 6 h, becoming statistically significant at 15 h; this effect was seen with both drugs. However, titers were similar to controls in groups 5. These results show that the interdigestive myoelectric complex is an important regulator at bacterial growth in the small intestine.

Action Potentials↗

[Antibiotic therapy in infectious endocarditis].

The most common causes of infective endocarditis, accounting for 65 to 85% of all cases, are viridans streptococci and other nonhemolytic streptococci. Enterococci are the offending microorganisms in 5 to 15%, staphylococci in 5 to 15% and gram-negative bacteria from the intestinal tract in 2 to 6%. In rare cases, infective endocarditis may be caused by any of a number of other pathogenic and nonpathogenic bacteria. Men over 60 years of age and women under 40 have a higher likelihood of contracting enterococcal endocarditis subsequent to febrile infections of the urogenital tract or after abortion; intravenous drug users tend to infections with gram-negative bacteria; patients with intravascular catheters who are administered cortisone, broad-spectrum antibiotics or cytostatic drugs are at risk of endocarditis from Candida or Aspergillus. At least two, but in general, five blood cultures should be drawn in short intervals. With the use of proper techniques for detection of aerobic and anaerobic microorganisms as well as fungi, positive blood cultures can be obtained in 95% of the patients. Antibiotics may be discontinued temporarily in pretreated patients. Bactericidal antibiotics are indicated. The following rule is valid as a guideline for adequate antibacterial chemotherapy: at maximal concentration after antibiotic administration, a bactericidal effect should still be demonstrated after 1:8 dilution of the patient's serum. Prior to receipt of blood culture findings, in forms tending to be subacute, treatment should be directed at streptococci and enterococci. If the course is more acute, in the presence of an intracardiac foreign body or in intravenous drug users, the antibiotic employed should also be effective against staphylococci.(ABSTRACT TRUNCATED AT 250 WORDS)

Adult↗

Protective immunity against Salmonella typhimurium elicited in mice by oral vaccination with phosphorylcholine encapsulated in poly(DL-lactide-co-glycolide) microspheres.

Encapsulation of vaccines in biodegradable microspheres provides excellent mucosal immunogens with a high potential for immunization against bacterial infections. We tested the protective immunity elicited by intragastric vaccination with phosphorylcholine (PC) encapsulated in poly(DL-lactide-co-glycolide) (DL-PLG) microspheres against Salmonella typhimurium in a mouse model of invasive intestinal infection. We chose PC as the antigen because it was found to elicit an immune response after intestinal exposure of mice to PC-bearing S. typhimurium and because anti-PC immunity protects mice against Streptococcus pneumoniae, another PC-bearing microorganism. Mice were primed intragastrically on days 1, 2, and 3 and boosted on days 28, 29, and 30 with PC (280 microg) coupled to porcine thyroglobulin (PC-thyr) encapsulated in DL-PLG microspheres, free PC-thyr, or blank microspheres. A significant rise in anti-PC immunoglobulin A (IgA) titers, as measured by an enzyme-linked immunosorbent assay, was observed in the intestinal secretions after immunization with PC-loaded microspheres, compared to the titers of mice immunized with free PC-thyr or blank microspheres. This antibody response peaked 14 days after the last boost and correlated with a highly significant resistance to oral challenge by S. typhimurium C5 (P < 10(-3)). Control mice were primed intraperitoneally on day 1 with 15 microg of PC in complete Freund's adjuvant and boosted on days 10, 14, and 20 with the same dose without adjuvant but via the same route. In these mice, the levels of anti-PC IgA in intestinal secretions were equivalent to those of the mice intragastrically immunized with PC-loaded microspheres, but protection was significantly weaker, suggesting that either the IgAs were not functional or that other immune mechanisms are important in protection. Taken together, our results highlight the potential of antigen encapsulation in DL-PLG microspheres for eliciting protective immunity against invasive intestinal bacterial diseases and suggest that a similar strategy could be used against diseases caused by other PC-bearing microorganisms.

Administration, Oral↗

Innate immunity of the gut: mucosal defense in health and disease.

The intestine is an important immune organ consisting of a complex cellular network, secreted peptides and proteins and other host defenses. Innate immunity plays a central role in intestinal immune defense against invading pathogens. It also serves as a bridge to the activation of the adaptive immune system. Pattern recognition molecules of microorganisms are an essential component for identifying invading pathogens. Toll-like receptors (TLRs), CARD15/NOD2 and scavenger receptors all serve as the pattern recognition receptors in the innate immune defense system. Secreted bactericidal peptides or defensins produced by the intestinal epithelia represent another crucial element of innate mucosal immune defense. Mutations in pattern recognition receptors and dysfunction of secretory bactericidal peptides may impair host immune defenses leading to an invasion of pathogens resulting in chronic inflammation of the gut. This review updates our current understanding of innate immunity of the gastrointestinal tract.

Humans↗

Modification of the intestinal microflora using probiotics and prebiotics.

Probiotics and prebiotics modulate the composition of the human gut microbiota. The beneficial effects may result from suppression of harmful microorganisms or stimulation of organisms which contribute in a positive way to the nutrition and health of the host. Both types of supplement represent an attempt to reconstitute the gut flora to its normal composition which has been adversely affected by dietary and environmental stresses.

Animals↗

[An analysis of the strain homogeneity of parietal biotopes of the gastrointestinal tract].

The lumen and parietal biotopes of the human gastrointestinal tract were investigated, i.e. the antibiotic sensitivity of strains E. coli and Enterococcus faecalis residing in them were comparatively analyzed. Certain isolation degrees not only of the parietal and fecal biotopes but also of certain micro-regions in the intestine wall were detected. The obtained data confirm a previously made assumption on the cluster distribution of microorganisms in the parietal mucin. The distinct strain differences between the parietal and fecal biotopes are not enough to completely extrapolate the available characteristics of the lumen microflora on the parietal microbiocenose. The above stated underlines the necessity in purpose-oriented research of parietal microbe populations of "internal" biotopes.

Adult↗

Resistance profile of Bacteroides fragilis isolated in Brazil. Do they shelter the cfiA gene?

The epidemiology of antimicrobial resistance of clinical isolates and human intestinal strains of Bacteroides fragilis has assumed great importance in the last few years since this microorganism, like other members of the B. fragilis group, can be responsible for the spread of resistance determinants. It is possible that the presence of B. fragilis in polluted aquatic environments might contribute to the spread of resistance. The antimicrobial resistance profile of 44 clinical B. fragilis strains isolated from 1981-1988 and 1991-1998 from the University hospital of Rio de Janeiro, and of 17 faecal and 17 polluted aquatic environmental B. fragilis strains isolated between 1991 and 1998 was determined. The susceptibility tests against penicillin, cefoxitin, imipenem, meropenem, clindamycin, chloramphenicol and metronidazole were performed by Etest in Wilkins-Chalgren agar enriched with 5% sheep blood. Motivated by some high MIC values for cefoxitin and meropenem, the cfiA gene, which codes for a metallo-beta-lactamase, was investigated among all strains, using PCR amplification. The resistance to penicillin was high in the samples from 1981 to 1988 (92.9%) and also in those from 1991 to 1998 (100%), although the MIC90 decreased from 256 mg/L to 24 mg/L. An increase in the resistance level to clindamycin and cefoxitin was seen from one decade to the other, the MIC90 values changing from 4 mg/L to 12 mg/L and from 8 mg/L to 32 mg/L, respectively. The susceptibility profile for metronidazole, chloramphenicol, imipenem and meropenem remained stable, although two clinical strains showed MICs of 6 mg/L and 8 mg/L against meropenem. Almost all human intestinal strains were resistant to penicillin and all of them were susceptible to imipenem, meropenem, chloramphenicol and metronidazole. The MICs of meropenem against two strains isolated from a polluted aquatic environment were 6 mg/L and 32 mg/L. The cfiA gene was detected in five strains, two of which were isolated from clinical specimens against which the MIC values of cefoxitin were high and three from an aquatic environment, whose susceptibility to both cefoxitin and meropenem ranged from sensitive to resistant.

Bacterial Proteins↗

[Studies On The Transmissibility Of Pathogenic-Organisms To Liver By Migrating Larvae Of Liver Fluke And Hookworm]

In order to confirm whether the migrating larvae of parasites could carry pathogenic organisms into liver and cause hepatitis, a series of experiments has been carried out. A. Clonorchis sinensis: Recovery rate of larvae in the abdominal cavity of rabbits: One to seven days after the administration adolescariae were recovered from the abdominal cavity in less than l % of the total number of metacercariae given. Generally, 1~6 larvae were found from each animal which was given 900~1,000 metacercariae, though many larvae were already found in the common bile ducts or remained still in intestine. Fate of Clonorchis sinensis in abdominal cavity: The young or mature worms which were introduced directly into the abdominal cavity were examined l5, 32, 40 and 42 days after the inoculation. Several larvae were found on the surface of liver in four animals. All the worms on the surface of the liver were dead and the biopsied liver tissues on the area where the worms were attached showed no pathological changes. Two of them were between bile duct and liver tissue but pus cell infiltration surrounding them was observed. In every case, pus cell infiltration was found in the peripheral portion of the liver and pus nodules on the surface of intestine and mesentery. The nodule in the intestinal wall contained the eggs of Clonorchis sinensis. Two worms in the abdominal cavity were still alive. From the above results it is suggested that the larvae of Clonorchis sinensis were capable of penetrating the intestinal wall and reaching the organs in the abdominal cavity and surviving for l5~42 days, but they were unable to penetrate the organs. No bacterial flora appeared from the lesion by culture method. Fate of Clonorchis sinensis which was inoculated into the peripheral region of liver: Small abscess was observed at the same area. Microscopically, the area became edematous and the vessels in the peripheral region were dilated. The parasites became necrotic and amorphous. Pathologically the lesions appeared as eosinophilic masses and neutrophile leukocytes were infiltrated surrounding the masses. In some cases, the dead worms were found apart from the original place of inoculation but no leukocyte infiltration was found. There was linear infiltration between the original site and the portion where the dead worm was found. The distance from the capsule varied from 0 to 4 mm. Sometimes, the eggs of Clonorchis sinensis were also found. In all cases, there were no living worms in liver tissues and hepatic ducts. In all case,. the bacteriological examination was negative. Do clonorchis sinensis transfer the microorganism? Five adult worms of clonorchis sinensis were incubated in the saline solution containing Staphylococcus aureus. The intestinal contents of these worms were cultured in the Nutient-agar plate and examined by Methylene Blue and Gram's stain. The area of liver tissue where the Clonorchis sinensis were inoculated showed no inflammatory changes after the 3 days of inoculation but no living Staphylococcus aureus was found in the culture media with which the pieces of liver tissues were smeared. B. Hookworm: Cutaneous infection: Four to eight days after the cutaneous infection of Ancylostoma caninum, the mice were sacrificed. Grossly, there was no abnormal finding in liver. The pieces of liver tissues were smeared on the Nutrient-agar plate, and cocci were found in four out of six examined. The microorganism were confirmed as the same species of Diplococcus pneumoniae which were grown in the hookworm culture media. Oral infection: 1,000 filariform larvae of Ancylostoma caninum were given orally. 24 hours later, the mice were sacrificed and the pieces of liver tissue were smeared on the Nutrient-agar plate. After 50 hours at 36 C, the bacterial colonies were examined bacteriologically. Staphylococcus albus was found from two out of four samples. Grossly there was no abnormality on the surface of liver, but microscopically there were spots like microabscesses which were infiltrated by leukocytes. The larvae were also found from other portions of liver tissues and they were surrounded by yellow colored material. In another experiment, a combination of Ancylostoma duodenale and Staphylococcus aureus was fed to mice. The mice sacrificed five days after the oral administration of Ancylostoma duodenale cultivated in the media containing Staphylococcus aureus. The liver pieces were examined routinely. The larvae cultivated in normal tap water which contained no Staphylococcus aureus was used as control. In the experimental mouse, the cocci appeared in the liver. Pathologically, microabscesses infiltrated with neutrophile leukocytes were found, but there was no manifestation of inflammatory change due to Staphylococcus aureus. There was only mechanical trauma due to the larvae penetration. Haemorrhage appeared only where the larvae were found.

Journal Article↗